Team:UC Chile/Cyanolux/Future

From 2012.igem.org

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<h1>Promoters Redesigned</h1>
<h1>Promoters Redesigned</h1>
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We are designing new versions of our chosen promoters. Now we have changed the promoter construction methodology in terms of the lenght of the upstream sequence considered.
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We [http://2012.igem.org/Team:UC_Chile/Results/LuxBrick#N-decanal_Effect_Characterization_-_.28BBa_K325909_and_BBa_K325905.29 have seen] that even if induced with substrate, our luciferase constructs don´t show luminescence, though the constructs´ sequences are verified and well characterized luciferases do show light emition.
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Our advisors in cyanobacteria have told us that sometimes promoter sequences can be placed several hundred of bases upstream the +1 site, even inside protein coding sequences.
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That´s why we are designing new versions of our chosen promoters. Now we have changed the promoter construction methodology in terms of the lenght of the upstream sequence considered.
Instead of considering just 150-200bp, up to 1000bp will be amplified from Synechocystis genome.
Instead of considering just 150-200bp, up to 1000bp will be amplified from Synechocystis genome.
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<h1>Transcriptional verification</h1>
<h1>Transcriptional verification</h1>
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Even if our promoters are o.k, colony PCRs can´t tell us wether the transcriptional machinery of our cyanobacteria is recognizing our constructs only  .
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Even if our promoters are o.k, colony PCRs can´t tell us wether the transcriptional machinery of our cyanobacteria is recognizing our constructs.
We are designig new primers to make  RT-PCRs that unmistakably verify transcription of the Lux genes  at the specified hours.
We are designig new primers to make  RT-PCRs that unmistakably verify transcription of the Lux genes  at the specified hours.

Revision as of 03:37, 27 September 2012

Project: Luxilla - Pontificia Universidad Católica de Chile, iGEM 2012